LOCUS       sequence-201396-        5590 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Cre and mRuby2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5590)
  AUTHORS   Meyer P, Maity P, Burkovski A, Schwab J, Mussel C, Singh K, Ferreira
            FF, Krug L, Maier HJ, Wlaschek M, Wirth T, Kestler HA, 
            Scharffetter-Kochanek K
  TITLE     A model of the onset of the senescence associated secretory 
            phenotype after DNA damage induced senescence.
  JOURNAL   PLoS Comput Biol. 2017 Dec 4;13(12):e1005741. doi: 
            10.1371/journal.pcbi.1005741. eCollection 2017 Dec.
  PUBMED    29206223
REFERENCE   2  (bases 1 to 5590)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS Comput
            Biol."; date: "2017-12-4"; volume: "13(12):e1005741. doi"; pages: " 
            10.1371/journal.pcbi.1005741. eCollection 2017 Dec"
COMMENT     Sequence Label: pCAG-Cre-T2A-mRuby2
FEATURES             Location/Qualifiers
     source          1..5590
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     158..180
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     242..261
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      289..298
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             295..1323
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MANLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             1333..1386
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1387..2097
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     primer_bind     complement(2186..2205)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2251..2306
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2305..2324)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2667..2683)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2667..2683)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2680..2702)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2691..2707
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2715..2721,2722..2739,2740..2745))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2760..2781
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2840..3035
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2886..3021
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3041..3175
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3078..3097)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3132..3151
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3243..3260)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3414..4002)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3494..3513)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4173..4964,4965..5033))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4796..4815
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5034..5138)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5169..5548
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
       61 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
      121 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
      181 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
      241 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcgc caccatggcc
      301 aatttactga ccgtacacca aaatttgcct gcattaccgg tcgatgcaac gagtgatgag
      361 gttcgcaaga acctgatgga catgttcagg gatcgccagg cgttttctga gcatacctgg
      421 aaaatgcttc tgtccgtttg ccggtcgtgg gcggcatggt gcaagttgaa taaccggaaa
      481 tggtttcccg cagaacctga agatgttcgc gattatcttc tatatcttca ggcgcgcggt
      541 ctggcagtaa aaactatcca gcaacatttg ggccagctaa acatgcttca tcgtcggtcc
      601 gggctgccac gaccaagtga cagcaatgct gtttcactgg ttatgcggcg gatccgaaaa
      661 gaaaacgttg atgccggtga acgtgcaaaa caggctctag cgttcgaacg cactgatttc
      721 gaccaggttc gttcactcat ggaaaatagc gatcgctgcc aggatatacg taatctggca
      781 tttctgggga ttgcttataa caccctgtta cgtatagccg aaattgccag gatcagggtt
      841 aaagatatct cacgtactga cggtgggaga atgttaatcc atattggcag aacgaaaacg
      901 ctggttagca ccgcaggtgt agagaaggca cttagcctgg gggtaactaa actggtcgag
      961 cgatggattt ccgtctctgg tgtagctgat gatccgaata actacctgtt ttgccgggtc
     1021 agaaaaaatg gtgttgccgc gccatctgcc accagccagc tatcaactcg cgccctggaa
     1081 gggatttttg aagcaactca tcgattgatt tacggcgcta aggatgactc tggtcagaga
     1141 tacctggcct ggtctggaca cagtgcccgt gtcggagccg cgcgagatat ggcccgcgct
     1201 ggagtttcaa taccggagat catgcaagct ggtggctgga ccaatgtaaa tattgtcatg
     1261 aactatatcc gtaacctgga tagtgaaaca ggggcaatgg tgcgcctgct ggaagatggc
     1321 gatgggagtg gagagggcag gggaagtctt ctaacatgcg gggacgtgga ggaaaatccc
     1381 ggccccatgg tgtctaaggg cgaagagctg atcaaggaaa atatgcgtat gaaggtggtc
     1441 atggaaggtt cggtcaacgg ccaccaattc aaatgcacag gtgaaggaga aggcaatccg
     1501 tacatgggaa ctcaaaccat gaggatcaaa gtcatcgagg gaggacccct gccatttgcc
     1561 tttgacattc ttgccacgtc gttcatgtat ggcagccgta cttttatcaa gtacccgaaa
     1621 ggcattcctg atttctttaa acagtccttt cctgagggtt ttacttggga aagagttacg
     1681 agatacgaag atggtggagt cgtcaccgtc atgcaggaca ccagccttga ggatggctgt
     1741 ctcgtttacc acgtccaagt cagaggggta aactttccct ccaatggtcc cgtgatgcag
     1801 aagaagacca agggttggga gcctaataca gagatgatgt atccagcaga tggtggtctg
     1861 aggggataca ctcatatggc actgaaagtt gatggtggtg gccatctgtc ttgctctttc
     1921 gtaacaactt acaggtcaaa aaagaccgtc gggaacatca agatgcccgg tatccatgcc
     1981 gttgatcacc gcctggaaag gttagaggaa agtgacaatg aaatgttcgt agtacaacgc
     2041 gaacacgcag ttgccaagtt cgccgggctt ggtggtggga tggacgagct gtacaagtga
     2101 gcggccgcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg
     2161 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca
     2221 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     2281 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     2341 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca
     2401 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt
     2461 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt
     2521 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc
     2581 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc
     2641 tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     2701 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     2761 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2821 ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc
     2881 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2941 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3001 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta
     3061 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3121 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg
     3181 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3241 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3301 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3361 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3421 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3481 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3541 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3601 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3661 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3721 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3781 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3841 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3901 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3961 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4021 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4081 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4141 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4201 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4261 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4321 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4381 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4441 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4501 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4561 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4621 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4681 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4741 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4801 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4861 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4921 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4981 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5041 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5101 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5161 cctgggtcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     5221 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     5281 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     5341 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     5401 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     5461 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     5521 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
     5581 ccccatctcc
//
